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Development of a small‐scale bioreactor: Application to in vivo NMR measurement
Author(s) -
Gmati Dorra,
Chen Jingkui,
Jolicoeur Mario
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20293
Subject(s) - bioreactor , in vivo , intracellular , chemistry , nuclear magnetic resonance spectroscopy , cytoplasm , biophysics , proton nmr , pi , chromatography , analytical chemistry (journal) , biochemistry , biology , stereochemistry , microbiology and biotechnology , organic chemistry
A perfused bioreactor allowing in vivo NMR measurement was developed and validated for Eschscholtzia californica cells. The bioreactor was made of a 10‐mm NMR tube. NMR measurement of the signal‐to‐noise ratio was optimized using a sedimented compact bed of cells that were retained in the bioreactor by a supporting filter. Liquid medium flow through the cell bed was characterized from a mass balance on oxygen and a dispersive hydrodynamic model. Cell bed oxygen demand for 4 h perfusion required a minimal medium flow rate of 0.8 mL/min. Residence time distribution assays at 0.8–2.6 mL/min suggest that the cells are subjected to a uniform nutrient environment along the cell bed. Cell integrity was maintained for all culture conditions since the release of intracellular esterases was not significant even after 4 h of perfusion. In vivo NMR was performed for 31 P NMR and the spectrum can be recorded after only 10 min of spectral accumulation (500 scans) with peaks identified as G‐6P, F‐6P, cytoplasmic Pi, vacuolar Pi, ATP γ and ADP β , ATP α and ADP α , NADP and NDPG, NDPG and ATP β . Cell viability was shown to be maintained as 31 P chemical shifts were constant with time for all the identified nuclei, thus suggesting constant intracellular pH. © 2004 Wiley Periodicals, Inc.